Pyridoxine and pyridoxal analogues: new uses

ABSTRACT

The invention provides pyridoxal and pyridoxine analogues, pharmaceutical compositions containing pyridoxine and pyridoxal analogues, and methods of administering pharmaceutical compositions containing a therapeutically effective amount of at least one of these analogues. In accordance with the present invention, the pyridoxal and pyridoxine analogues can be used in the treatment or prevention of of heparin induced thrombocytopenia (HIT, stroke, and ischemia, and in the treatment of symptoms thereof. The the pyridoxal and pyridoxine analogues can be used in neuroprotection.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority from U.S. provisionalapplication No. 60/216,907 filed Jul. 7, 2000, and is acontinuation-in-part of U.S. Pat. No. 6,417,204 filed Jul. 6, 2001, andis a continuation-in-part of U.S. patent application Ser. No. 10/147,263filed May 15, 2002, which is pending.

FIELD OF THE INVENTION

[0002] The present invention relates to pyridoxine and pyridoxalanalogue compounds, pharmaceutical compositions containing thepyridoxine and pyridoxal analogue compounds, and methods of treatmentusing a therapeutically effective amount of the pyridoxine and pyridoxalanalogue compounds. The pyridoxine or pyridoxal analogues can be used inthe treatment of undesired stroke, ischemia, and heparin inducedthrombocytopenia (HIT), or related diseases and symptoms thereof, aswell as providing neuroprotection.

BACKGROUND OF THE INVENTION

[0003] Pyridoxal-5′-phosphate (PLP), an end product of vitamin B₆metabolism, plays a vital role in mammalian health. Vitamin B₆ typicallyrefers to pyridoxine, which is chemically known as2-methyl-3-hydroxy-4,5-di(hydroxymethyl)pyridine and is represented byformula I:

[0004] Yet two additional compounds, pyridoxal of formula II

[0005] and pyridoxamine of formula III

[0006] are also referred to as vitamin B₆. All three compounds serve asprecursors to pyridoxal-5′-phosphate (PLP), which is chemically known as3-hydroxy-2-methyl-5-[(phosphonooxy) methyl]-4-pyridine-carboxaldehydeand is represented by formula IV:

[0007] PLP is tho biologically active form of vitamin B₆ inside cellsand in blood plasma. Mammals cannot synthesize PLP de novo and must relyon dietary sources of the precursors pyridoxine, pyridoxal, andpyridoxamine, which are metabolized to PLP. For instance, mammalsproduce PLP by phosphorylating pyridoxine by action of pyridoxal kinaseand then oxidizing the phosphorylated product.

[0008] PLP is a regulator of biological processes and a cofactor in morethan one hundred enzymatic reactions. It has been shown to be anantagonist of a purinergic receptor, thereby affecting ATP binding; ithas been implicated in modulation of platelet aggregation, it is aninhibitor of certain phosphatase enzymes; and it has been implicated inthe control of gene transcription. In previous patents (U.S. Pat. No.6,051,587 and U.S. Pat. No. 6,043,259) the role ofpyridoxal-5′-phosphate, and its precursors pyridoxal and pyridoxine(vitamin B₆), in mediating cardiovascular health and in treatingcardiovascular related diseases is disclosed. PLP is also a coenzyme incertain enzyme-catalyzed processes, for example, in glycogenolysis atthe glycogen phosphorylase level, in the malate asparatate shuttleinvolving glycolysis and glycogenolysis at the transamination level, andin homocysteine metabolism.

[0009] There is a need to identify and administer drugs that can mimicone or more of the known biological actions of vitamin B-6 congeners butthat are more potent than the vitamin B-6 congeners in their specificmode of action

SUMMARY OF THE INVENTION

[0010] The present invention provides for pyridoxine and pyridoxalanalogues, pharmaceutical compositions containing the pyridoxine andpyridoxal analogues, and methods for treatment based on administrationof therapeutically effective amounts of the pyridoxine and pyridoxalanalogues. Compounds and compositions of the invention can be used forthe treatment of cardiovascular or related diseases and symptomsthereof.

[0011] The invention provides pyridoxine and pyridoxal analogues ofFormula V:

[0012] or a pharmaceutically acceptable acid addition salt addition saltthereof, wherein:

[0013] R₅ is CH₂OH or CHO;

[0014] R₁ is

[0015] n is an integer of 1 to 5;

[0016] R₂, R₃, and R₄ are each independently

[0017] hydrogen;

[0018] alkyl;

[0019] aryl or biaryl,

[0020] wherein the aryl or biaryl can be substituted with a cyano,alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy;

[0021] amino;

[0022] acylamino;

[0023] anilino,

[0024] wherein the aniline ring can be substituted with a cyano, alkyl,alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy,

[0025] nitro; or

[0026] guanidino.

[0027] In another aspect, the invention is directed to a pharmaceuticalcomposition that includes a pharmaceutically acceptable carrier incombination with a therapeutically effective amount of a compound ofFormula V or a pharmaceutically acceptable acid addition salt of acompound of Formula V.

[0028] In another aspect, the invention is directed to a method oftreating cardiovascular or related diseases and symptoms thereof. Themethod includes administering to a mammal a therapeuticaly effectiveamount of a compound of Formula V or a pharmaceutically acceptable acidaddition salt of a compound of Formula V in a unit dose form. The methodcan further include concurrent administration of another therapeuticagent.

[0029] In another aspect, the invention is directed to a method ofreducing platelet aggregation and the symptoms there of a mammalianpatient. The method includes administering to the mammalian patient atherapeutically effective amount of a compound of Formula V. In someinstances the patient may suffer from heparin-induced thrombocytopenia(HIT).

[0030] In another aspect, the invention is directed to a method ofreducing or preventing heparin-induced thrombus formation in a mammalianpatient suffering from HIT. The method includes administering to themammalian patient a therapeutically effective amount of a compound ofFormula V.

[0031] In another aspect, the invention is directed to a method ofreducing or preventing brain damage ill a mammalian patient sufferingfrom stroke or ischemia. The method includes administering to themammalian patient a therapeutically effective amount of a compound ofFormula V.

[0032] In another aspect, the invention is directed to a method ofreducing brain damage in a mammalian patient by neuroprotection. Themethod includes administering to the mammalian patient a therapeuticallyeffective amount of a compound of Formula V.

DETAILED DESCRIPTION OF THE INVENTION

[0033] The invention provides pyridoxal and pyridoxine analogues andpharmaceutical compositions containing these pyridoxine and pyridoxalanalogues. The pyridoxine and pyridoxal analogues can be used in theTreatment of cardiovascular or related diseases and symptoms thereof.

[0034] Cardiovascular or related diseases include, for example, cerebralischemia, cerebral hemorrhage, ischemic stroke, hemorrhagic stroke,hypertension, myocardial infarction, ischemia reperfusion injury,myocardial ischemia, congestive heart failure, blood coagulationdisorders, cardiac hypertrophy, and platelet aggregation. Cardiovascularor related diseases also includes diseases that arises from thromboticand prothrombotic states in which the coagulation cascade is activatedsuch as, for example, deep vein thrombosis, disseminated intravascularcoagulopathy, and pulmonary embolism.

[0035] Heart failure is a pathophysiological condition in which theheart is unable to pump blood at a rate commensurate with therequirement of the metabolizing tissues or can do so only from anelevated filling pressure (increased load). Thus, the heart has adiminished ability to keep up with its workload. Over time, thiscondition leads to excess fluid accumulation, such as peripheral edema,and is referred to as congestive heart failure.

[0036] When an excessive pressure or volume load is imposed on aventricle, myocardial hypertrophy (i.e., enlargement of the heartmuscle) develops as a compensatory mechanism. Hypertrophy permits theventricle to sustain an increased load because the heart muscle cancontract with greater force. However, a ventricle subjected to anabnormally elevated load for a prolonged period eventually fails tosustain an increased load despite the presence of ventricularhypertrophy, and pump failure can ultimately occur.

[0037] Heart failure can arise from any disease that affects the heartand interferes with circulation. For example, a disease that increasesthe heart muscle's workload, such as hypertension, will eventuallyweaken the force of the heart's contraction, Hypertension is a conditionin which there is an increase in resistance to blood flow through thevascular system. This resistance leads to increases in systolicpressure, diastolic blood pressure, or both. Hypertension placesincreased tension on the left ventricular myocardium, causing it tostiffen and hypertrophy, and accelerates the development ofatherosclerosis in the coronary arteries The combination of increaseddemand and lessened supply increases the likelihood of myocardialischemia leading to myocardial infarction, sudden death, arrhythmias,and congestive heart failure.

[0038] Ischemia is a condition in which an organ or a part of the bodyfails to receive a sufficient blood supply. When an organ is deprived ofa blood supply, it is said to be hypoxic. An organ will become hypoxiceven when the blood supply temporarily ceases, such as during a surgicalprocedure or during temporary artery blockage. Ischemia initially leadsto a decrease in or loss of contractile activity. When the organaffected is the heart, this condition is known as myocardial ischemia,and myocardial ischemia initially leads to abnormal electrical activity.This can generate an arrhythmia. When myocardial ischemia is ofsufficient severity and duration, cell injury can progress to celldeath—i.e., myocardial infarction—and subsequently to heart failure,hypertrophy, or congestive heart failure.

[0039] Ischemic reperfusion of the organ occurs when blood flow resumesto an organ after temporary cessation. For example, reperfusion of anischemic myocardium can counter the effects of coronary occlusion, acondition that leads to myocardial ischemia. Ischemic reperfusion to themyocardium can lead to reperfusion arrhythmia or reperfusion injury. Theseverity of reperfusion injury is affected by numerous factors, such as,for example, duration of ischemia, severity of ischemia, and speed ofreperfusion. Conditions observed with ischemia reperfusion injuryinclude neutrophil infiltration, necrosis, and apoptosis.

[0040] Heparin-induced thrombocytopenia (“HIT”) is an immune-mediatedsyndrome. HIT is caused by antibodies (HIT-Ig) which typically recognizea complex of platelet factor 4 and heparin The formation of theantibody/antigen complex activates platelets and endothelial cells andgenerates procoagulent microparticles. Clinical features of HIT callinclude thrombocytopenia, thrombosis, and heparin-induced skin lesions.(Warkentin, Ann, Rev. Med., 50: 129-147 (1999)).

[0041] HIT is characterized by an abrupt reduction in platelet countfollowing treatment of the patient with heparin. It can also cause theformation of clots on subsequent treatment with heparin. Previousattempts to identify compounds effective in preventing this subsequentimmune-induced (heparin-induced) clotting have not proven entirelysatisfactory.

[0042] The exact dose and route of administration of the compound ofFormula V will vary based on the patient's size, condition, gender, andmedical history. In some instances it will be desirable to administer acompound of Formula V to achieve a plasma concentration of between about1 M and 1000 μM. In some instances a plasma concentration of betweenabout 300 μM and 700 μM will be desirable. In some instances a plasmaconcentration of between about 450 μM and 550 μM will be desirable.

[0043] In some instances, it may be desirable to administer compound Vintravenously at a dose of between about 2 to 100 mg/kg body weight. Insome instances this dose will be administered between one and four timesdaily to maintain a desired plasma concentration of the compound.

[0044] The compounds of the invention may be administered to induce aplasma level of said compound of between about 100 nM and 10,000 μM.They may be administered to a therapeutically effective dose betweenabout 0.1 to 100 mg/kg body weight per day. The may be administered to adosage between 10 mg/kg and 100 mg/kg body weight. They may beadministered to a dosage is between 20 mg/kg and 80 mg/kg body weight.

[0045] The compounds of the invention may be administered intravenously,orally, sublingually, intraperitoneally, transdermally, intramuscularlyor subcutaneously. In a preferred embodiment, they may be administeredintravenously. In a further preferred embodiment, they may beadministered orally.

[0046] Pyridoxal and Pyridoxine Analogue Compounds

[0047] The invention provides pyridoxal and pyridoxine analoguecompounds of Formula V:

[0048] or pharmaceutically acceptable acid addition salts thereof,wherein:

[0049] R₅ is CH₂OH or CHO;

[0050] R₁ is

[0051] n is an integer of 1 to 5;

[0052] R₂, R₃, and R₄ are each independently

[0053] hydrogen;

[0054] alkyl;

[0055] aryl or biaryl;

[0056] wherein the aryl or biaryl can be substituted with a cyano,alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy;

[0057] amino;

[0058] acylamino;

[0059] anilino,

[0060] wherein the aniline ring can be substituted with a cyano, alkyl,alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy;

[0061] nitro; or

[0062] guanidino.

[0063] As used herein, the term “alkyl” refers to a straight or branchedsaturated aliphatic hydrocarbon chain having 1 to 8 carbon atoms, suchas, for example, methyl, ethyl, propyl, isopropyl (1-methylethyl),butyl, tert-butyl (1,1-dimethylethyl), and the like. The alkyl chain canbe interrupted by a heteroatom, such as, for example, a nitrogen,sulfur, or oxygen atom, forming an alkylaminoalkyl, alkylthioalkyl, oralkoxyalkyl. Examples of alkyl chain interrupted by a heteroatomsinclude methylaminoethyl, ethylthiopropyl, methoxymethyl, and the like.The alkyl can be substituted at the terminal carbon by groups such ashydroxy, alkoxy, alkanoyloxy, alkoxycarbonyl, or carboxy.

[0064] The term “alkoxy” refers to an alkyl group joined to an oxygenatom. In some embodiments, the alkoxy has 1 to 4 carbon atoms in astraight or branched chain, such as, for example, methoxy, ethoxy,propoxy, isopropoxy (1-methylethoxy), butoxy, tert-butoxy(1,1-dimethylethoxy), and the like.

[0065] As used herein, the term “alkanoyloxy” refers to a group offormula

[0066] Examples of an alkanoyloxy include methanoyloxy, ethanoyloxy,propanoyloxy, and the like.

[0067] The term “halo” refers to a bromo, chloro, or fluoro group. Insome embodiments, the halo is fluoro.

[0068] Pharmaceutically acceptable acid addition salts of the compoundsof Formula V include salts derived from nontoxic inorganic acids such ashydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic,hydrofluoric, phosphorous, and the like, as well as the salts derivedfrom nontoxic organic acids, such as aliphatic mono- and dicarboxylicacids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids,alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonicacids, etc. Such salts thus include sulfate, pyrosulfate, bisulfate,sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate,dihydiogenphosphate, metaphosphate, pyrophosphate, chloride, bromide,iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate,oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate,mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate,phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate,lactate, malcate, tartrate, methanesulfonate, and the like. Alsocontemplated are salts of amino acids such as arginate and the like andgluconate, galacturonate, n-methyl glutamine, etc. (see, e.g., Berge etal., J. Pharmaceutical Science, 66: 1-19 (1977).

[0069] The acid addition salts of the basic compounds arc prepared bycontacting the free base form with a sufficient amount of the desiredacid to produce the salt in the conventional manner. The free base formmay be regenerated by contacting the salt form with a base and isolatingthe free base in the conventional manner. The free base forms differfrom their respective salt forms somewhat in certain physical propertiessuch as solubility in polar solvents, but otherwise the salts areequivalent to their respective free base for purposes of the presentinvention.

[0070] In one embodiment of Formula V, R₁ is

[0071] Preferably, R₂ is hydrogen, alkyl, or amino.

[0072] In another embodiment of Formula V, R₁ is

[0073] Preferably, R₂ and R₃ are each independently hydrogen, alkyl,amino, or nitro.

[0074] In another embodiment of Formula V, R₁ is

[0075] Preferably, R₂, R₃, and R₄ are independently hydrogen, alkyl, oramino. In a particularly preferred embodiment, R₂ is hydrogen, R₃ ismethyl, and R₄ is hydrogen.

[0076] In another embodiment of Formula V, R₁ is

[0077] In still another embodiment of Formula V, R₁ is

[0078] In a particularly preferred embodiment, R₁ is

[0079] Methods of Preparing Pyridoxal and Pyridoxine Analogue Compounds

[0080] Another aspect of the invention provides a method for preparingthe pyridoxine and pyridoxal analogues, The compounds of the inventioncan be prepared from a compound of Formula VI, VII, VIII, IX, or X:

R₆═(CH₂)_(p)OH where p=1 to 5   VI

R₆═(CH₂)_(q)Br where q=1 to 5   VII

R₆═(CH₂)_(r)CHO where r=0 to 4   VIII

R₆═(CH₂)₅N₃ where s=1 to 5   IX

R₆═(CH₂)_(t)NH₂ where t=1 to 5   X

[0081] A compound of Formula VI, VII, VIII, IX, or X can be used to formthe compounds of Formula V through a series of chemical reactions toproduce pyridoxine analogues. The pyridoxine analogues can besubsequently oxidized to produce the corresponding pyridoxal analogues.

[0082] In some embodiments of the invention, the pyridoxine andpyridoxal analogues are formed by reacting a bromide compound of FormulaVII with a substituted or unsubstituted tetrazole, a substituted orunsubstituted triazole, or a substituted or unsubstituted imidazole. Thetetrazole, triazole, or imidazole can be substituted with an aryl,biaryl, amino, acylamino, anilino, or guanidine. An aryl or biaryl canbe further substituted with a cyano, alkyl, alkoxy, amino, hydroxy,halo, nitro, or alkanoyloxy group An aniline can be further substitutedwith a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxygroup.

[0083] For example, as shown in Scheme 1, a bromide compound of FormulaVII (q=1) can be reacted with 1H-tetrazole to produce derivative XI. Thederivative XI is then treated with acetic acid to produce5-tetrazolepyridoxine XII. Oxidation of 5-tetrazolepyridoxine XII in thepresence of a catalyst such as manganese dioxide can be use to producethe corresponding pyridoxal XIII.

[0084] In other embodiments of the invention, an aldehyde of FormulaVIII is formed by reacting an alcohol of Formula VI with a suitableoxidizing agent such as maganese dioxide. The pyridoxine and pyridoxalanalogues of Formula V are formed by reacting an aldehyde of FormulaVIII with a substituted or unsubstituted triazole, a substituted orunsubstituted imidazole, or a substituted or unsubstituted alanine. Thetriazole, or imidazole can be substituted with an aryl, biaryl, amino,acylamino, anilino, or guanidine. An aryl or biaryl can be furthersubstituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, oralkanoyloxy group. An aniline can be further substituted with a cyano,alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy group.

[0085] For example, an aldehyde of Formula VIII (r=0) can react with2-methylinidazoline to form the protected imidazoline derivative XXVIIIaccording to Scheme 2. The protected imidazoline derivative XXVII can behydrolyzed to the imidazoline XXIX.

[0086] In another example, a compound of the invention can be preparedby reacting an aldehyde of Formula VIII (r=0) with 4-cyano aniline asshown in Scheme 3 to form a Schiff base. The Schiff base XXXII isreacted with a strong reducing agent such as, for example, sodiumborohydride. The resulting amine XXXIII can react with ethanol in thepresence of dry hydrogen chloride gas to form a compound of FormulaXXXIV. The compound of Formula XXXIV can be treated with 2 M NH₃ in MeOHin a pressurized vessel to form the compound of Formula XXXV.

[0087] In another embodiment of preparing the compounds of Formula V, anamine of Formula X can react with a substituted guanidine.

[0088] For example, as shown in Scheme 4, an amine compound of Formula X(s=1) can react with a protected guanidine compound to give theguanidine derivative XXIII The protection groups can be removed withtrifluoro acetic acid to form a compound of Formula XXV.

[0089] In another embodiment of preparing the compounds of Formula V, anazide of Formula IX is the precursor. The isopropylidene group ishydrolyzed initially. The hydroxy groups formed from the hydrolysisreaction are then protected by reacting with a reagent such as, forexample, tert-butyldimethylsilyl chloride. The azide group can behydrogenated to an amine group. The resulting amine compound can reactwith an aromatic group containing a substituted or unsubstituted aryl orbiaryl isocyanate or a substituted thioisocyanate. The aryl or biarylcan be substituted wit a cyano, alkyl, alkoxy, amino, hydroxy, halo,nitro, or alkanoyloxy. The protection groups added after the hydrolysisreaction can be removed to form a pyridoxine compound.

[0090] For example, as shown in Scheme 5, an arylurea and arylthioureasubstituted compound of the invention can be prepared using an azidecompound of Formula IX (s=1). The isopropylidene group can be hydrolyzedby treatment with acetic acid to form compound of Formula XLI. Theunprotected hydroxyl groups are reacted with tert-butyldimethylsilylchloride to form XLII. The azide group can be hydrogenated with hydrogenin the presence of a catalyst to form the amine XLIII. The amine canreact with 4-fluorophenylisocyanate giving compound XLIV. The synthesesof compounds XLVI and XLVII followed the procedure outlined for XLIV andXLV using 4-fluorophenylthioisocyanate in place of4-fluorophenylisocyanate.

[0091] The products of the reactions described herein are isolated byconventional means such as extraction, distillation, chromatography, andthe like.

[0092] One skilled in the art can recognize other variations in thereaction sequences and in the appropriate reaction conditions from theanalogous reactions shown or otherwise known that may be appropriatelyused in the above-described processes to make the compounds of Formula Vherein.

[0093] Pharmaceutical Compositions

[0094] Although it is possible for a pyridoxine and pyridoxal analoguecompound of the invention to be administered alone in a unit dosageform, the compounds are typically administered in admixture as apharmaceutical composition to provide a unit dosage form. The inventionprovides pharmaceutical compositions containing at least one pyridoxineor pyridoxal analogues compound of Formula V. A pharmaceuticalcomposition comprises a pharmaceutically acceptable carrier incombination with a compound of Formula V or a pharmaceuticallyacceptable acid addition salt of a compound of Formula V.

[0095] A pharmaceutically acceptable carrier includes, but is notlimited to, physiological saline, ringers, phosphate-buffered saline,and other carriers known in the art. Pharmaceutical compositions canalso include additives such as, for example, stabilizers, antioxidants,colorants, excipients, binders, thickeners, dispersing agents,reabsorption enhancers, buffers, surfactants, preservatives,emulsifiers, isotonizing, agents, and diluents. Pharmaceuticallyacceptable carriers and additives are chosen such that side effects fromthe pharmaceutical compound are minimized and the performance of thecompound is not canceled or inhibited to such an extent that treatmentis ineffective.

[0096] Methods of preparing pharmaceutical compositions containing apharmaceutically acceptable carrier in combination with a therapeuticcompound of Formula V or a pharmaceutically acceptable acid additionsalt of a compound of Formula V are known to those of skill in the art.All methods can include the step of bringing the compound of theinvention in association with the carrier and additives, Theformulations generally are prepared by uniformly and intimately bringingthe compound of the invention into association with a liquid carrier ora finely divided solid carrier or both, and then, if necessary, shapingthe product into the desired unit dosage forms.

[0097] For oral administration as a suspension, the compositions can beprepared according to techniques well known in the art of pharmaceuticalformulation. The compositions can contain microcrystalline cellulose forimparting bulk, alginic acid or sodium alginate as a suspending agent,methylcellulose as a viscosity enhancer, and sweeteners or flavoringagents. As immediate release tablets, the compositions can containmicrocrystalline cellulose, starch, magnesium stearate and lactose orother excipients, binders, extenders, disintegrants, diluents andlubricants known in the art.

[0098] For administration by inhalation or aerosol, the compositions canbe prepared according to techniques well known in the art ofpharmaceutical formulation. The compositions can be prepared assolutions in saline, using benzyl alcohol or other suitablepreservatives, absorption promoters to enhance bioavailability,fluorocarbons or other solubilizing or dispersing agents known in theart.

[0099] For administration as injectable solutions or suspensions, thecompositions can be formulated according to techniques well-known in theart, using suitable dispersing or wetting and suspending agents, such assterile oils, including synthetic mono- or diglycerides, and fattyacids, including oleic acid.

[0100] For rectal administration as suppositories, the compositions canbe prepared by mixing with a suitable non-irritating excipient, such ascocoa butter, synthetic glyceride esters or polyethylene glycols, whichare solid at ambient temperatures, but liquefy or dissolve in the rectalcavity to release the drug.

[0101] Method of Treatment Using Pyridoxal and Pyridoxine AnalogueCompounds

[0102] In another aspect of the invention, methods are provided for thetreatment of cardiovascular or related diseases and symptoms thereof.

[0103] As used herein the terms “treatment” and “treating” as usedherein include preventing, inhibiting, alleviating and healing vitaminB₆ cardiovascular or related diseases or symptoms thereof Treatment canbe carried out by administering a therapeutically effective amount of acompound of the invention A “therapeutically effective amount” as usedherein includes a prophylactic amount, for example, an amount effectivefor preventing or protecting against the above mentioned diseases orsymptoms thereof; or an amount effective for alleviating or healing theabove mentioned diseases or symptoms thereof.

[0104] A physician or veterinarian of ordinary skill readily determinesa mammalian subject who is exhibiting symptoms of any one or more of thediseases described above. Regardless of the route of administrationselected, a compound of Formula V or a pharmaceutically acceptable acidaddition salt of a compound of Formula V can be formulated intopharmaceutically acceptable unit dosage forms by conventional methodsknown in the pharmaceutical art. An effective but nontoxic quantity ofthe compound is employed in treatment The compounds can be administeredin enteral unit dosage forms, such as, for example, tablets,sustained-release tablets, enteric coated tablets, capsules,sustained-release capsules, enteric coated capsules, pills, powders,granules, solutions, and the like. They can also be administeredparenterally, such as, for example, subcutaneously, intramuscularly,intradermally, intramammarally, intravenously, and by otheradministrative methods known in the art.

[0105] The ordinarily skilled physician or veterinarian will readilydetermine and prescribe the therapeutically effective amount of thecompound to treat the disease for which treatment is administered. In soproceeding, the physician or veterinarian could employ relatively lowdosages at first, subsequently increasing the dose until a maximumresponse is obtained. Typically, the particular disease, the severity ofthe disease, the compound to be administered, the route ofadministration, and the characteristics of the mammal to be treated, forexample, age, sex, and weight, are considered in determining theeffective amount to administer. Administering a therapeutic amount of acompound of the invention for treating cardiovascular or relateddiseases or symptoms thereof, is in a range of about 0.1-100 mg/kg of apatient's body weight, more preferably in the range of about 0.5-50mg/kg of a patient's body weight, per daily dose. The compound can beadministered for periods of short and long duration. Although someindividual situations can warrant to the contrary, short-termadministration, for example, 30 days or less, of doses larger than 25mg(kg of a patient's body weight is preferred to long,-termadministration. When long-term administration, for example, months oryears, is required, the suggested dose usually does not exceed 25 mg/kgof a patient's body weight.

[0106] A therapeutically effective amount of a compound of Formula V ora pharmaceutically acceptable addition salt of a compound of Formula Vfor treating the above-identified diseases or symptoms thereof can beadministered prior to, concurrently with, or after the onset of thedisease or symptom. A compound of the invention can be administeredconcurrently. “Concurrent administration” and “concurrentlyadministering” as used herein includes administering a compound of theinvention and another therapeutic agent in admixture, such as, forexample, in a pharmaceutical composition or in solution, or separately,such as, for examples separate pharmaceutical compositions or solutionsadministered consecutively, simultaneously, or at different times butnot so distant in time such that the compound of the invention and theother therapeutic agent cannot interact and a lower dosage amount of theactive ingredient cannot be administered.

[0107] In one embodiment of the invention, a method is provided fortreating cardiovascular or related diseases comprising administering toa mammal a therapeutically effective amount of a compound of Formula Vor a pharmaceutically acceptable addition salt of a compound of FormulaV in a unit dosage form. The cardiovascular or related diseases that canbe treated include hypertrophy, hypertension, congestive heart failure,heart failure subsequent to myocardial infarction, myocardial ischemia,cerebral ischemia, ischemia reperfusion injury, arrhythmia, myocardialinfarction, blood coagulation, or platelet aggregation. Preferably, thecardiovascular disease treated is hypertrophy, congestive heart failure,arrhythmia, or ischemia reperfusion injury.

[0108] The compound of the invention can also be administered to treatcardiovascular diseases and other diseases that arise from thromboticand prothrombotic states in which the coagulation cascade is activated,such as, for example, deep vein thrombosis, disseminated intravascularcoagulopathy, Kasabach-Merritt syndrome, pulmonary embolism, myocardialinfarction, stroke, thromboembolic complications of surgery, andperipheral arterial occlusion. A compound of the invention may also beuseful in the treatment of adult respiratory distress syndrome, septicshock, septicemia, or inflammatory responses, such as edema and acute orchronic atherosclerosis, because thrombin has been shown to activate alarge number of cells outside of the coagulation process, such as, forexample, neutrophils, fibroblasts, endothelial cells, and smooth musclecells.

[0109] The method for treating cardiovascular or related diseases canfurther comprise concurrent administration of other therapeutic agentsalready known to be suitable for treating the above-identified diseases.For example, methods of the invention include concurrently administeringa compound of Formula V or a pharmaceutically acceptable acid additionsalt of a compound of Formula V in combination with a therapeuticcardiovascular compound to treat hypertrophy, hypertension, congestiveheart failure, heart failure subsequent to myocardial infarction,myocardial ischemia, ischemia reperfusion injury, arrhythmia, ormyocardial infarction. Preferably, the cardiovascular disease treated ishypertrophy, congestive heart failure, arrhythmia, or ischemiareperfusion injury,

[0110] Other therapeutic cardiovascular compounds that can beconcurrently administered with a compound or composition of theinvention include an angiotensin converting enzyme inhibitor, anangiotensin II receptor antagonist, a calcium channel blocker, anantithrombolytic agent, a β-adrenergic receptor antagonist, avasodilator, a diuretic, an α-adrenergic receptor antagonist, anantioxidant, and a mixture thereof. In one embodiment, a compound of theinvention is administered concurrently with PPADS (pyridoxalphosphate-6-azophenyl-2′,4′-disulphonic acid), also a therapeuticcardiovascular compound, or concurrently with PPADS and another knowntherapeutic cardiovascular compound as already described.

[0111] Preferably the other therapeutic cardiovascular compound, whichis concurrently administered with a compound of Formula V or apharmaceutically acceptable acid addition salt of a compound of FormulaV, is an angiotensin converting enzyme inhibitor, an angiotensin IIreceptor antagonist, a diuretic, an α-adrenergic receptor antagonist, ora calcium channel blocker.

[0112] Known angiotensin converting enzyme inhibitors include, forexample, captopril, enalapril lisinopril, benazapril, fosinopril,quinapril, ramipril, spirapril, imidapril, and moexipril.

[0113] Examples of known angiotensin II receptor antagonists includeboth angiotensin I receptor subtype antagonists and angiotensin IIreceptor subtype antagonists. Suitable angiotensin II receptorantagonists include losartan and valsartan.

[0114] Suitable calcium channel blockers include, for example,verapamil, diltiazem, nicardipine, nifedipine, amlodipine, felodipine,nicardipine, and bepridil.

[0115] Examples of known β-adrenergic receptor antagonists includeatenolol, propranolol, timolol, and metoprolol.

[0116] Suitable vasodilators include, for example, hydralazine,nitroglycerin, and isosorbide dinitrate.

[0117] Suitable diurctics include, for example, furosemide, diuril,amiloride, aid hydrodiuril.

[0118] Suitable α-adrenergic receptor antagonists include, for example,prazosin, doxazocin, and labetalol.

[0119] Suitable atioxidants include vitamin E, vitamin C, andisoflavones.

[0120] These other therapeutic cardiovascular compounds are generallyused to treat cardiovascular or related diseases as well as symptomsthereof. A skilled physician or veterinarian readily determines asubject who is exhibiting symptoms of any one or more of the diseasesdescribed above and makes the determination about which compound isgenerally suitable for treating specific cardiovascular conditions andsymptoms.

[0121] For example, myocardial ischemia can be treated by theadministration of a compound of Formula V or a pharmaceuticallyacceptable acid addition salt of a compound of Formula V concurrentlywith another therapeutic agent. Other suitable therapeutic agentsinclude, for example, a angiotensin converting enzyme inhibitor, anangiotensin II receptor antagonist, a calcium channel blocker, anantithrombolytic agent, a β-adrenergic receptor antagonist, a diuretic,an I-adrenergic receptor antagonist, or a mixture thereof

[0122] As another example, congestive heart failure can be treated bythe administration of a compound of Formula V or a pharmaceuticallyacceptable acid addition salt of a compound of Formula V concurrentlywith another therapeutic agent. Other suitable therapeutic agentsinclude, for example, a angiotensin converting enzyme inhibitor, anangiotensin II receptor antagonist, a calcium channel blocker, avasodilator, a diuretic, or a mixture thereof.

[0123] Myocardial infarction can be treated by the administration of acompound of Formula V or a pharmaceutically acceptable acid additionsalt of a compound of Formula V concurrently with another therapeuticagent. Other suitable therapeutic agents include, for example, aangiotensin convert enzyme inhibitor, a calcium channel blocker, anantithrombolytic agent, a β-adrenergic receptor antagonist, a diuretic,an α-adrenergic receptor antagonist, or a mixture thereof.

[0124] Hypertension can be treated by the administration of a compoundof Formula V or a pharmaceutically acceptable acid addition salt of acompound of Formula V concurrently with another therapeutic agent. Othersuitable therapeutic agents include, for example, an angiotensinconverting enzyme inhibitor, a calcium channel blocker, a β-adrenergicreceptor antagonist, a vasodilator, a diuretic, an α-adrenergic receptorantagonist, or a mixture thereof.

[0125] Arrhythmia can be treated by the administration of a compound ofFormula V or a pharmaceutically acceptable acid addition salt of acompound of Formula V concurrently with another therapeutic agent. Othersuitable therapeutic agents include, for example, a calcium channelblocker, a β-adrenergic receptor antagonist, or a mixture thereof.

[0126] Blood clots in the arteries can be reduced or removed by theadministration of a compound of Formula V or a pharmaceuticallyacceptable acid addition salt of a compound of Formula V concurrentlywith an antithrombolytic agent. Antithrombolytic agents known in the artinclude antiplatelet agents, aspirin, and heparin.

[0127] Hypertrophy can be treated by the administration of a compound ofFormula V or a pharmaceutically acceptable acid addition salt of acompound of Formula V concurrently with another therapeutic agent. Othersuitable therapeutic agents include, for example, an angiotensinconverting enzyme inhibitor, an angiotensin II receptor antagonist, acalcium channel blocker, or a mixture thereof.

[0128] Ischemia reperfusion injury can be treated by the administrationof a compound of Formula V or a pharmaceutically acceptable acidaddition salt of a compound of Formula V concurrently with anothertherapeutic agent. Other suitable therapeutic agents include, forexample, an angiotensin converting enzyme inhibitor, an angiotensin IIreceptor antagonist, a calcium channel blocker, or a mixture thereof.

[0129] Platelet aggregation can be treated by the administration of acompound of Formula V or a pharmaceutically acceptable acid additionsalt of a compound of Formula V concurrently with another therapeuticagent. Other suitable therapeutic agents include, for example,Danaparoid; Lepirudin; Hirudin; Argatroban; and in some instances,Heparin and/or Warfarin; or a mixture thereof.

[0130] This invention is further characterized by the followingexamples. These examples are not meant to limit the scope of theinvention but are provided for exemplary purposes to more fully describethe invention. Variation within the scope of the invention will beapparent to those skilled in the art.

EXAMPLES

[0131] Preparation of Starting Materials

[0132] Bromide VII (q=1) was prepared by a literature procedure;Imperalli et at, J. Org. Chem., 60, 1891-1894 (1995). Alcohol VI (p=1)was prepared by bubbling HCl gas into a solution of pyridoxinehydrochloride (50 g, 0.24 mol) in acetone (500 mL) at 0-5° C. (ice bath)until the solution became clear. Diethyl ether (ca. 1 L) was added toinduce precipitation of the hydrochloride salt which was filtered off.The salt was dissolved in a mixture of methylene chloride (ca. 1 L) andsaturated aqueous NaHCO₃ (ca. 500 mL). The layers were separated and theorganic layer washed with saturated aqueous NaHCO₃. The combined organiclayers were dried (MgSO₄), and evaporated to give 40.5 g (80%) of acolourless solid.

[0133] Alcohol VI (p=1) was dissolved in dichloromethane and cooled to0° C. Small of amounts of triphenylphosphine and N-bromosuccinimide wereadded alternately over a period of about five minute. The reactionmixture was stirred for about 20 minutes and then concentrated in vacuo.The crude product, bromide VII (q=1) was purified by flashchromatography using a 2:1 mixture of ether and hexanes as the eluent.The product was used immediately.

[0134] Aldehyde VIII (r=0) was identified by comparison to data in theliterature; Kortynk et al., J. Org. Chem., 29, 574-579 (1964). MnO₂(Aldrich 21,764-6) (49.9 g, 85%, 487 mmol) was added to a solution ofalcohol VI (p=1) (25 g, 119.6 mmol) in toluene (900 ml). The resultingmixture was stirred at 40° C. for 24 hours then filtered through Celite.The mother liquor was evaporated to give a light yellow solid. The solidwas recrystallized from hexane: ethyl ether (1:1) to give a light yellowsolid. The solid was filtered and washed with hexane: ethyl ether (1:1)to give the pure aldehyde VIII (17.51 g, 71%).

[0135] Azide IX (s=1) and amine X (t=1) were prepared from bromide VII(q=1). Bromide VII (q=1) (1.08 g. 4.0 mmol) in anhydrous DMF (20 ml) wastreated with sodium azide (260 mg, 4.0 mmol) at room temperature. Afterone hour stirring at room temperature, the solution was extracted withdiethyl ether (5×20 mL). The combined extracts were washed with water(10 mL) and brine (10 mL) followed by drying (MgSO₄). The solvent wasevaporated and the crude product was purified by chromatography onsilica gel using ethyl ether: hexanes (2:1) as eluent to give the azideIX (s=1) as a colourless liquid (552 mg, 60%). ¹H NMR (CDCl₃, TMS) δ1.57(s, 6H), 2.42 (s, 3H), 4.23 (s, 2H), 4.86 (s, 2H), 7.96 (s, 1H).

[0136] The purified azide IX (s=1) (100 mg, 0.4 mmol) was dissolved in95% ethanol and hydrogenated at 1 atm in presence of Lindlar catalyst(50 mg) for one hour. The catalyst was removed by filtration (Celite),and the solvent removed to give the crude amine X (t=1). Purification bychromatography on silica gel using CH₂Cl₂:MeOH (5:1) as eluent gave theproduct (80 mg, 82% ) ¹HNMR (CD₂Cl₂) 1.53 (s, 6H), 2.34 (s, 3H), 3.72(s, 2H), 4.91 (s, 2H), 5.31 (s, 2H), 7.93 (s, 1H.

[0137] All other reagents used in the following examples can bepurchased from Aldrich Chemical Company (Milwaukee, Wis. or Allentown,Pa.).

Example 1

[0138] Synthesis of Tetrazole Substituted Pyridoxine Analogue of FormulaXII

[0139] A mixture of tetrazole (94.2 mg, 1.29 mmol) and pulverizedanhydrous potassium carbonate (1.5 g) in anhydrous acetonitrile (10 mL)was stirred at 0° C. for 15 minutes. The bromide VII (q=1) (350 mg, 1.29mmol) in anhydrous acetonitrile (3 mL) was then added to the reactionmixture maintaining the reaction temperature for the next 30 Minutes.After the completion of the reaction, routine work-up gave the crudeproduct. Purification of the crude mixture on silica gel column gave thedesired product XI in appreciable yields.

[0140]¹H nmr (CDCl₃, TMS): δ1:52 (6H, s) 2.44 (3H, s), 4.77 (2H, s),5.47 (2H, s), 8.07 (1H, s), 8.55 (1H, s, -tetrazole-II).

[0141] The purified derivative XI (100 mg, 0.4 mmol) was then taken in80% aqaeous acetic acid (10 mL) and heated at 60° C. for 1 hour.Purification by chromatography on silica gel gave the5-tetrazolepyridoxine XII in good yields.

[0142]¹H nmr (CD₃OD, TMS): δ2.42 (3H, s), 4.96 (2H, s), 5.97 (2H, s),7.92 (1H, s), 8.69 (1H, s, terazole H).

Example 2

[0143] Synthesis of Tetrazole Substituted Pyridoxal Analogue of FormulaXIII

[0144] The 5-tetrazole pyridoxine XII (100 mg, 0.42 mmol) was dissolvedin anhydrous toluene (10 mL). To the solution was added activatedmanganese dioxide (243 mg, 2.76 mmol), and the reaction mixture heatedat 40° C. for 2 hours to ensure complete oxidation. Filtration of thecatalyst, followed by evaporation of the solvent gave the crude residuewhich was easily purified by chromatography on silica gel to give thedesired aldehyde XIII in 70% yield.

[0145]¹H nmr (CD₂Cl₂, TMS): δ2.82 (3H, s), 6.00 (1H, s), 6.15 (1H, s),8.11 (1H, s), 8.57 (1H, s, tetrazole-H), 10.77 (1H, s, aldeydic-H).

Example 3

[0146] Synthesis of Tetrazole Substituted Pyridoxine Analogue of FormulaXV

[0147] A mixture of tetrazole (94.2 mg, 1.29 mmol) and pulverizedanhydrous potassium carbonate (1.5 g) in anhydrous acetonitrile (10 mL)was stirred at 0° C. for 15 minutes. The bromide VII (q=1) (350 mg, 1.29mmol) in anhydrous acetolnitrile (3 mL) was then added to the reactionmixture which was maintained at 0° C. for the next 30 minutes. Routinework-up gave the crude product. Purification of the crude mixture bychromatography on silica gel gave the desired product XIV in appreciableyields.

[0148]¹H nmr (CDCl₃, TMS ): δ1:53 (6H, s), 2.42 (3H, s), 4.91 (2H, s),5.66 (2H,s), 8.14 (1H, s), 8.50 (1H, s,-tetrazole-H).

[0149] The purified derivative XIV (100 mg, 0.4 mmol) was dissolved in80% aqueous acetic acid (10 mL) and heated at 60° C. for 1 hour.Purification by chromatography on silica gel column gave the 5-tetrazolepyridoxine XV in good yields.

[0150]¹H nmr (CD₃OD, TMS): δ2.43 ( 3H, s), 4.89 (2H, s), 5.77 (2H, s),7.91 (1H,s), 9.17 (1H, s, terazole H).

Example 4

[0151] Synthesis of Tetrazole Substituted Pyridoxine Analogue of FormulaXVII

[0152] A mixture of aminotetrazole (110.2 mg, 1.30 mmol) and pulverizedanhydrous potassium carbonate (1.5 g) in anhydrous acetonitrile (10 mL)was stirred at 0° C. for 15 minutes. The bromide VII (q=1) (360 mg, 1.30mmol) in anhydrous acetonitrile (3 mL) was added to the reaction minutewhich was maintained at 0° C. for the next 30 minutes. Routine workupgave the crude product. Purification of the crude mixture bychromatography on a silica gel column gave the desired product XVI inappreciable yields.

[0153]¹H nmr(CD₃OD, TMS): δ1.52 (6H, s), 2.36 (3H, s), 4.96 (2H, s),5.56 (2H, s), 7.96 (1H, s).

[0154] The purified derivative XVI (100 mg, 0.37 mmol) was dissolved in80% aqueous acetic acid (10 mL) and heated at 60° C. for 1 hour.Purification by chromatography on silica gel gave the 5-tetrazolepyridoxine XVII in good yields ¹H nmr (CD₃OD, TMS): δ2.42 (3H,s), 4.94(2H, s), 5.66 (2H, s), 7.91 (1H, s), 7.87 (1H, s).

Example 5

[0155] Synthesis of Triazole Substituted Pyridoxine Analogue of FormulaXIXa

[0156] A mixture of triazole (136 mg, 2.00 mmol) and pulverizedanhydrous potassium carbonate (2.5 g) in anhydrous acetonitrile (20 mL)was stirred at 0° C. for 15 minutes. The bromide VII (q=1) (720 mg, 2.00mmol) in anhydrous acetonitrile (5 mL) was added to the reaction mixturewhich was maintained at 0° C. for the next 30 minutes. Routine work-upgave the crude product. Purification of the crude mixture bychromatography on a silica gel column gave the desired product XVIIIa inappreciable yield.

[0157]¹H nmr (CDCl₃, TMS): δ1:53 (6H, s), 2.42 (3H, s), 4.80 (2H s),5.24 (2H, s), 7.94 (1H,s,-triazole-H), 7.99 (1H, s, -triazole-H), 8.15(1H, s).

[0158] The purified derivative XVIIIa (100 mg, 0.35 mmol) was dissolvedin 80% aqueous acetic acid (10 mL) and heated at 60° C. Purification bychromatography on silica gel column gave the 5-triazole pyridoxine XIXain good yield.

[0159]¹H nmr (CD₃OD, TMS ): δ2.42 (3H, s), 4.91 (2H, s), 5.50 (2H, s),7.82 (1H, s,-triazole-H), 7.97 (1H, s, -triazole-H), 8.52 (1H, s).

Example 6

[0160] Synthesis of Triazole Substituted Pyridoxal Analogue of FormulaXXa

[0161] Following Scheme 8, 5-triazole pyridoxine XIXa (100 mg, 0.42mmol) was dissolved in anhydrous toluene (10 mL). To the solution wasadded activated manganese dioxide (243 mg, 2.76 mmol), and the reactionmixture heated at 40° C. for 2 hours to ensure complete oxidation.Filtration of the catalyst, followed by evaporation of the solvent gavea crude residue that was easily purified by chromatography on silica gelto give the desired aldehyde XXa in 70% yield.

[0162]¹H nmr (CD₃OD, TMS): δ2.70 (3H, s), 6.00 (2H, s), 6.28 (2H, s),8,41 (1H, s,-triazole-H), 8.85 (1H, s, -triazole-M), 9.85 (1H, s).

Example 7

[0163] Synthesis of Triazole Substituted Pyridoxine Analogue of FormulaXIXb

[0164] Following Scheme 8, a mixture of 3-aminotriazole (142 mg, 2.00mmol) and pulverized anhydrous potassium carbonate (2.5 g) in anhydrousacetonitrile (20 mL) was stirred at 0° C. for 15 minutes. The bromideVII (q=1) (720 mg, 2.00 mmol) in anhydrous acetonitrile (5 mL) was addedto the reaction mixture which was maintained at 0° C. for the next 30minutes. Routine workup gave the crude product. Purification of thecrude mixture by chromatography on silica gel the desired product XVIIIbin appreciable yields.

[0165]¹H nmr (CD₃OD, TMS): δ1.57 (6H, s), 2.36 (3H, s), 4.84 (2H, s),4.99 (2H, s), 7.84 (1H, s, -triazole-H, 8.13 (1H, s).

[0166] The purified derivative XVIIIb (100 mg, 0.35 mmol) was dissolvedin 80% aqueous acetic acid (10 mL) and heated at 60° C. for 1 hour.Purification by chromatography on silica gel gave the 3-aminotriazolepyridoxine XIXb in good yields.

[0167]¹H nmr (DMSO-d₆, TMS): δ2.34 (3H, s), 4.72 (2H, s), 5.17 (2H, s),7.79 (1H, s, -triazole-H), 8.03 (1H, S).

Example 8

[0168] Synthesis of Triazole Substituted Pyridoxine Analogue of FormulaXIXc

[0169] Following Scheme 8, a mixture of 5-aminotriazole (142 mg, 2.00mmol) and pulverized anhydrous potassium carbonate (2.5 g) in anhydrousacetonitrile (20 mL) was stirred at 0° C. for 15 minutes. The bromideVII (q=1) (720 mg, 2.00 mmol) in anhydrous acetonitrile (5 mL) was addedto the reaction mixture which was maintained 0° C. for the next 30minutes. Routine work-up gave the crude product. Purification of thecrude mixture by chromatography on silica gel gave the desired productXVIIIc in appreciable yields.

[0170]¹H nmr (CD₃OD, TMS): δ1.53 (6H, s), 2.36 ( 3H, s), 4.84 (2H, s),5.04 (2H, s), 7.46 (1H, s, -triazole-H), 7.68 (1H, s).

[0171] The purified derivative XVIIIc (100 ng, 0.35 mmol) was dissolvedin 80% aqueous acetic acid (10 mL) and heated at 60° C. for 1 hour.Purification by chromatography on silica gel gave the 3-aminotriazolepyridoxine XIXc in good yield.

[0172]¹H nmr (DMSO-d₆, TMS): δ2.32 (3H, s), 4.75 (2H, s), 5.11 (2H, s),7.35 (1H, S, -triazole-H), 7.58 (1H, s).

Example 9

[0173] Synthesis of Imidazole Substituted Pyridoxine Analogue of FormulaXXIIa

[0174] A mixture of 2-methylimidazol (164 mg, 2.00 mmol) and pulverizedanhydrous potassium carbonate (2.5 g) in anhydrous acetonitrile (20 mL)was stirred at 0° C. for 15 minutes. The bromide VII (q=1) (720 mg, 2.00mmol) in anhydrous acetonitrile (5 mL) was added to the reaction mixturewhich was maintained at 0° C. for the next 30 minutes. Routine work-upgave the crude product. Purification of the crude mixture bychromatography on silica gel gave the desired product XXIa inappreciable yields.

[0175]¹H nmr (CDCl₃, TMS): δ1.51 (6H, s), 2.40 ( 3H, s), 2.42 (3H, s,imidazole-CH₃), 4.50 (2H, s), 4.88 (2H, s), 6.65 (1H, s, imidazole-H),6.93 (1H, 6, imidazole-H), 7.82 (1H, s).

[0176] The purified derivative XXIa (100 mg, 0.35 mmol) was thendissolved in 80% aqueous acetic acid (10 mL) and heated at 60° C. for 1hour. Purification by chromatography on silica gel gave the2-methylimidazole pyridoxine XXIIa in good yield.

[0177]¹H nmr (DMSO-d₆, TMS): δ2.40 (3H, s), 2.21 (3H, s, imidazole-CH₃),5.11 (2H, s), 5.20 (2H, s), 6.77 (1H, s, imidazole-H), 6.92 (1H, s,imidazole-H), (2H, s), 7.47 (1H, s).

Example 10

[0178] Synthesis of Imidazole Substituted Pyridoxine Analogue of FormulaXXIIb

[0179] Following Scheme 9, a mixture of 4-methylimidazole (164 mg, 2.00mmol) and pulverized anhydrous potassium carbonate (2.5 g) in anhydrousacetonitrile (20 mL) was stirred at 0° C. for 15 minutes. The bromideVII (q=1) (720 mg, 2.00 mmol) in anhydrous acetonitrile (5 mL) was addedto the reaction mixture which was maintained at 0° C. for the next 30minutes. Routine work-up gave the crude product. Purification of thecrude mixture by chromatography on silica gel gave the desired productXXIb in appreciable yields.

[0180]¹H nmr (CDCl₃, TMS): δ1.50 (6H, s), 2.43 ( 3H, s), 2.20 (3H s,imidazole-CH₃), 4.54 (2H, s), 4.92 (2H, s), 6.52 (1H, s, imidazole-H),7.42 (1H, s, imidazole-H), 7.94 (1H, s).

[0181] The purified derivative XXIb (100 mg, 0.35 mmol) was dissolved in80% aqueous acetic acid (10 mL) and heated at 60° C. for 1 hour.Purification by chromatography on silica gel gave the 4-methylimidazolepyridoxine XXIb in good yield.

[0182]¹H nmr (CDCl₃, TMS): δ2.20 (3H, s, imidazole-CH₃), 2.46 (3H, s),4.72 (2H, s), 4.90 (2H, s), 6.48 (1H, s, imidazole-H), 7.24 (1H, s,imidazole-H), 7.84 (1H, s).

Example 11

[0183] Synthesis of Imidazole Substituted Pyridoxine Analogue of FormulaXXIIc

[0184] Following Scheme 9, a mixture of 4-nitroimidazole (172 mg, 2.00mmol) and pulverized anhydrous potassium carbonate (2.5 g) in anhydrousacetonitrile (20 mL) was stirred at 0° C. for 15 minutes. The bromideVII (q=1) (720 mg, 2.00 mmol) in anhydrous acetonitrile (5 mL) was addedto the reaction mixture which was maintained at 0° C. for the next 30minutes. Routine work-up gave the crude product Purification of thecrude mixture by chromatography on silica gel gave the desired productXXIc in appreciable yield.

[0185]¹H nmr (CDCl₃, TMS): δ1.52 (6H, s), 2.42 (3H, s), 4.60 (2H, s),5.09 (2H, s), 7.46 (1H, s, imidazole-H), 7.69 (1H, s, imidazole-H), 8.01(1H, s).

[0186] The purified derivative XXIc (110 mg, 0.35 mmol) was dissolved in80% aqueous acetic acid (10 mL) and heated at 60° C. for 1 hour.Purification by chromatography on silica gel gave the 4-nitroimidazolepyridoxine XXIIc in good yield.

[0187]¹H nmr (DMSO-d₆, TMS): δ2.42 (3H, s), 4.74 (2H, s), 5.35 (2H, s),7.77 (1H, S, imidazole-H), 7.95 (1H, s), 8.14 (1H, s, imidazole-H).

Example 12

[0188] Synthesis of Imidazole Substituted Pyridoxine Analogue of FormulaXXIId

[0189] Following Scheme 9, a mixture of 2-aminoimidazole (132 mg, 2.00mmol) and pulverized anhydrous potassium carbonate (2.5 g) in anhydrousacetonitrile (20 mL) was stirred at 0° C. for 15 minutes. The bromideVII (q=1) (720 mg, 2.00 mmol) in anhydrous acetonitrile (5 mL) was addedto the reaction mixture which was maintained at 0° C. for the next 30minutes, Routine work-up gave the crude product. Purification of thecrude mixture by chromatography on silica gel gave the desired productXXId in appreciable yield.

[0190]¹H nmr (CD₃OD, TMS): δ1.52 (6H, s), 2.35 (3H, s), 4.68 (2H, s),4.88 (2H, s), 6.44 (1H, s, imidazole-H), 6.53 (1H, s, imidazole-H), 7.64(1H, s).

[0191] The purified derivative XXId (106 mg, 0.35 mmol) was thendissolved in 80% aqueous acetic acid (10 mL) and heated at 60° C. for 1hour. Purification by chromatography on silica gel gave the,2-aminoimidazole pyridoxine XXIId in good yield.

[0192]¹H nmr (CD₃OD, TMS): δ2.15(3H, s), 4.55 (2H, s), 4.75 (2H, s),6.24 (1H, s, imidazole-H), 6.32 (1H, s, imidazole-H), 7.23 (1H, s).

Example 13

[0193] Synthesis of Guanidine Substituted Pyridoxine Analogue of FormulaXXV

[0194] A solution of 5-aminopyridoxine derivative X (t=1) (90 mg, 0.43mmol) in anhydrous dichloromethane (5 mL), was added a solution oftriflated BOC-guanidine derivative (1553.6 mg, 0.39 mmol) in anhydrousdichloromethane (5 mL) and anhydrous triethylamine (60 μL). Thissolution was stirred at room temperature for one hour. The reactionmixture was washed with 2M sodium bisulfite (10 mL), followed bysaturated aqueous NaHCO₃. Evaporation of the dichloromethane left aresidue that was purified by chromatography on silica gel to give theguanidine derivative XXIII in appreciable yield.

[0195]¹H nmr (CD₂Cl₂, TMS ): δ1.46 (9H, s), 1.48 (9H, s), 1.54 (6H, s),4.40 (2H, s), 4.88 (2H, s), 7.95 (1H, s), 8.43 (NH, s), 11.48 (NH, s).

[0196] The purified derivative XXIII (100 mg, 0.36 mmol) was thendissolved in 20% trifluoroacetic acid in anhydrous dichloromethane (10mL) and stirred at room temperature for 1 hour. Purification of thereaction mixture gave the two products XXIV and XXV.

[0197]¹H nmr of XXIV (MeOD, TMS): δ1.64 (6H, s), 2.63 (3H, s), 4.55 (2H,s), 5.10 (2H, s), 7.52 (1H, m), 8.13 (1H, s).

[0198]¹H nmr of XXV (DMSO-d₆, TMS): δ2.54 (3H, s), 3.96 (1H, s), 4.54(2H, d), 4.81 (2H, s), 5.96 (1H, br s, NH), 7.44 (3H, br, NH), 8.07 (1H,s).

Example 14

[0199] Synthesis of Aminotriazole Substituted Pyridoxal Analogue ofFormula XXVIII

[0200] A pyridoxal derivative VIII (r=0) (400 mg, 1.91 mmol) and3-aminotriazole (178 mg, 2.12 mmol) in anhydrous toluene (20 mL), washeated in a three neck flask, fitted with a condenser and a Dean Starktrap, at 100° C. for 24 hours. Routine workup gave the crude product,which was then purified by chromatography on silica gel to give theprotected triazoline derivative XVI in modest yields.

[0201]¹H nmr (CD₃OD, TMS): δ1.58 (6H, s), 2.46 (3H, s), 3.31(1H, s),5.31 (2H, s), 8.41 (1H, s), 9.23 (1H, s, -triazoline CH).

[0202] The fully protected pyridoxine derivative XXVI (206 mg, 0.88mmol) was dissolved in 80% aqueous acetic acid (10 mL) and heated at 60°C. for 1 hour. Purification by chromatography on silica gel gave thetriazoline substituted pyridoxine XXVII in good yield.

[0203]¹H nmr (CD₃OD, TMS: δ2.46 (3H, s), 5.10 (2H, m), 6.78 (1H, s,-triazoline CH), 7.92 (1H, s).

Example 15

[0204] Synthesis of Imidazoline Substituted Pyridoxine Analogue ofFormula XXIX

[0205] A pyridoxal derivative VIII (r=0) (2.07 g, 10.00 mmol) and2-methylimidazoline (1.68 g, 20.00 mmol) in anhydrous toluene (50 mL),was heated in a three neck flask fitted with a condenser and a DeanStark trap, at 100° C. for 24 hours. Routine workup gave the crudeproduct, which was then purified by chromatography on silica gel to givethe protected imidazoline derivative XXVIII in modest yields.

[0206]¹H nmr (CD₃OD, TMS): δ1.56 (6H, s), 2.38 (3H, s), 3.76 (4H, s),4.98 (2H, s), 6.63 (1H, s, vinylic CH), 7.26 (1H, s, vinylic-CH), 8.20(1H, s).

[0207] The fully protected pyridoxine derivative XXVIII (100 mg, 0.43mmol) was dissolved in 80% aqueous acetic acid (5 mL) and heated at 60°C. 1 hour. Purification by chromatography on silica gel gave theimidazoline XXIX in good yield.

[0208]¹H nmr (CD₃OD, TMS): δ2.14 (3H, s), 3.09 (2H, s), 3.56 (4H, s),4.74 (1H, s, vinylic CH), 4.87 (1H, s, vinylic-CH), 5.24 (1H, d, NH) 726(1H, s).

Example 16

[0209] Synthesis of Imidazole Substituted Pyridoxine Analogue of FormulaXXXI

[0210] Pyridoxal derivative VIII (r=0) (800 mg, 3.82 mmol) and2-aminoimidazole (346 mg, 4.2 mmol) in anhydrous dimethyl sulfoxide (75mL), was heated in a three neck flak, fitted with a condenser and a DeanStark trap, at 100° C. for three days. Routine workup gave the crudeproduct, which was then purified by chromatography on silica gel to givethe protected cyclic guanidine derivative XXX in modest yield.

[0211]¹H nmr (MeOD, TMS): δ1.58 (6H, 5) 2.49 (3Hs, H) 5.26 (2H, s), 7.07(2H, s imidazole-H), 8.49 (1H. s), 9.27 (1H, s-vinylic H).

[0212] The protected pyridoxine derivative XXX (110 mg, 0.44 mmol) wasdissolved in 80% aqueous acetic acid (5 mL) and heated at 60° C. for 1hour. Purification by chromatography on silica gel gave the cyclicguanidine XXXI in good yield.

[0213]¹H nmr (MeOD, TMS): δ2.45 (3H, s), 4.96(2H, m), 5.12 (2H, dd),6.43 (1H, d), 7.92 (1H, s).

Example 17

[0214] Synthesis of Aminophenylamidine Substituted Pyridoxine Analogueof Formula X

[0215] In a 250 mL three-necked flak fitted with a condenser and a Deanstark tap was added protected pyridoxine aldehyde VIII (r=0) (3.31 g, 16mmol). 4-cyano aniline (1.89 g, 15.9 mmol), p-toluenesulfonic acid (0.3g, 1.6 mmol) and dry benzene (60 mL). The reaction mixture was thenheated to reflux for 16 hours. The reaction mixture was then washed with2N NaOH (5 mL) followed by brine (10 mL) and the organic, layer driedwith anhydrous Na₂SO₄. Removal of the solvent gave the crude product,which was purified by silica gel column chromatography, using the eluant100:1/CH₂Cl₂:2M NH₃-MeOH. XXXII was obtained as a pure yellow solid.(2.98 g, 61% yield).

[0216]¹H NMR (CDCl₃) δ1.58 (s 6H) 2.49 (s, 3H) 5.27 (s, 2H) 7.20 (d, 2H)7.68 (d, 2H) 8.31 (s, 1H) 8.44 (s, 1H).

[0217] The Schiff base XXXII (358 mg, 1.0 mmol) was dissolved in HOAc (5mL) and the solution cooled to 0° C. Sodium borohydride (57 mg, 1.5mmol) was added in portions, while stirring continued for 10 minutes a0° C., and then at room temperature for another 10 minutes. The reactionwas quenched by adding 5 N NaOH (1.8 mL) to bring the pH of the solutionto 9. The product was then extracted with diethyl ether (2×10 mL) anddried over anhydrous Na₂SO₄. Evaporation of the solvent, followed bypurification on silica gel column chromatography in EtOAc; Hexanes/1:1gave XXXIII as a pale yellow solid (300 mg, 90%)

[0218]¹HNMR (CDCl₃) δ1.54 (s, 6H), 2.40 (s, 3H) 4.20 (s, 2H) 4.36 (br,1H) 4.84 (s, 2H) 6.62 (d, 2H) 7.45 (d, 2H) 8.00 (s, 1H).

[0219] Into a solution of the amine XXXIII (81 mg, 0.3 mmol) in ethanol(6 mL) at 0° C. was bubbled hydrogen chloride gas (dry) for 30 min. Thereaction mixture was slowly allowed to reach the room temperature andthen stirred at this temperature for 16 hours. The solution was againcooled to 0° C., and then degassed by bubbling N₂ through it for 2 h. Onevaporation of the solvent XXXIV was obtained as a light yellow solid.

[0220]¹HNMR (MeOD) δ1.57 (t, 3H) 2.64 (s, 3H) 4.54 (q, 2H) 4.65 (s, 2H)5.14 (s, 2H) 6.82 (d, 2H) 7.89 (d, 2H) 8.00 (s, 1H).

[0221] In a sealed high pressure flask containing the crude compoundXXXIV (106 mg, 0.3 mmol) was added 2 M NH₃-MeOH (10 mL) and the mixturecooled to −78° C., and stirred at this temperature for 15 minutes. Thereaction mixture was gradually warmed to rt, and then heated to 80° C.for 2 h. The reaction mixture was again cooled to −78° C. and then thesealed flask was opened. The solution was then transferred to around-bottomed flask and the solvent evaporated to dryness. Purificationon silica gel column using 100:20:1/CH₂Cl₂; MeOH: H2O as eluent gaveXXXV as white solid (88 mg, 82%).

[0222]¹HNMR (MeOD) δ2.43 (s, 3H) 4.45 (s, 2H) 4.96 (s, 2H) 6.78 (d, 2H)7.64 (d, 2H) 7.84 (s, 1H).

Example 18

[0223] Synthesis of Aminophenylamidine Substituted Pyridoxine Analogueof Formula XL.

[0224] The procedure for the preparation of XXXVI was similar to thatused for the synthesis of compound XXXIIII. The crude product, XXXVI,was an auburn solid and was used in the next step without purification.

[0225]¹HNMR (CDCl₃) δ159 (s, 6H) 2.49 (s, 3H) 5.30 (s, 2H) 7.06 (d, 2H)7.51 (d, 2H) 8.30 (s, 1H) 8.47 (s, 1H).

[0226] The procedure for the preparation of XXXVII was similar to thatused for the synthesis of compound XXXIII. The crude product, XXXVII,was purified by silica gel column, using 5:1:1.5/CH₂Cl₂:MeOH:H₂O, aseluant to give a yield white solid. (66% yield)

[0227]¹HNMR (CDCl₃) δ1.55 (s, 6H) 2.41 (s, 3H) 3.68 (br, 1H) 4.12 (d,2H) 4.87 (s, 2H) 6.51 (d, 2H) 7.27 (d, 2H) 8.01 (s, 1H).

[0228] Compound XXXVII (665 mg, 1.83 mmol) was dissolved in diglyme (15mL), followed by the addition of Pd (PPh₃)₄ (63 mg, 0.05 mmol). Themixture was stirred for 10 min, and then p-cyanophenylboronic acid (269m, 2.01 mmol) was added to the reaction mixture followed by sodiumbicarbonate (461 mg in 8 mL H₂O, 5.49 mmol). The reaction mixture washeated to 95° C. in an oil bath for 5 min, and then stirred at roomtemperature for 1.5 h. On evaporation of the solvent a dark purple crudeproduct, XXXVIII, was obtained which was purified by silica gel columnchromatography (1:1/BtOAc: Haxane, 32% yield.).

[0229]¹HNMR (CDCl₃) δ1.56 (s, 6H) 2.42 (s, 3H) 3.96 (br, 1H) 4.22 (d,2H) 4.91 (s, 2H) 6.72 (d, 2H) 7.47 (d, 2H) 7.65 (q, 4H) 8.06 (s, 1H).

[0230] The syntheses of compound XXXIX and XL were accomplished by theprocedures outlined for compounds XXXIV and XXXV. Overall yield of XLwas 63% for the two steps.

[0231]¹HNMR (XXXIX) (MeOD) δ1.62 (t, 3H) 2.62 (s, 3H) 4.56 (s, 2H) 4.63(q, 2H) 5.17 (s, 2H) 6.78 (d, 2H) 7.60 (d, 2H) 7.83 (d, 2H) 8.02 (s, 1H)8.05 (d, 2H).

[0232]¹HNMR (XL) (MeOD) δ2.43 (s, 3H) 4.39 (S, 2H) 4.98 (s, 2H) 6.76 (d,2H) 7.54 (d, 2H) 7.80 (m, 4H) 7.89 (s, 1H).

Example 19

[0233] Synthesis of Arylurea and Arylthiourea Substituted PyridoxineAnalogues of Formulas XLIII and XLIV

[0234] The azide IX (t=1) (790 mg, 3.4 mmol) was dissolved in 80%aqueaul HOAc (40 mL) and heated at 60° C. for 16 hours, to hydrolyze theisopropylidene group. Co-distillation with toluene to remove acetic acidgave the crude product, which was purified by silica gel columnchromatography. (1:1→4:1/EtOAc: Hexane), giving XLI as a white solid(410 mg, 63% yield.)

[0235]¹HNMR, (MeOD) δ2.43 (s, 3H) 4.42 (s, 2H) 4.92 (s, 2H) 7.86 (s,1H).

[0236] To the de-blocked azide XLI (388 mg, 2.0 mmol) in dry DMF (10 mL)was added imidazole (545 mg, 8.8 mmol) and TBDPSiCl (1.2 ml, 4.4 mmol).The reaction mixture was then heated to 50° C. and kept stirring forovernight. The reaction was then cooled to room temperature, and thenextracted with diethyl ether (2×25 mL), followed by washing the etherextract by water (2×10 mL) and brine (1×10 mL). The ethereal solutionwas then dried over MgSO₄ and solvent evaporated to dryness to give XLIIas a white solid. (1.3 g, 97% yield).

[0237]¹H NMR (CDCl₃) δ0.87 (s, 9H) 0.96 (s, 9H) 2.15 (s, 3H) 4.49 (s,2H) 4.52 (s, 2H) 7.38 (m, 20H) 8.03 (s, 1H).

[0238] To the compound XLII (1.3 g. 1.94 mmol) dissolved in methanol (20mL) was added Lindlar catalyst (600 mg) and the mixture hydrogenatedunder a stream H₂ for 1.5 hours to complete the reaction. The routinework up and purification gave the amine XLIII (0.95 g 761%).

[0239]¹H NMR(CDCl₃) δ0.83 (s, 9H) 0.96 (s, 9H) 1.51 (br, 2H) 2.11 (s,3H) 3.89 (s, 2H) 4.57 (s, 2H) 7.33 (m, 20) 8.09 (s, 1H).

[0240] A mixture of XLIII (645 mg 1.0 mmol) and the4-fluorophenylisocyanate (0.12 mL, 1.0 mmol) in dry toluene (10 mL) wasrefluxed for 16 hours. After the removal of the solvent, the crudeproduct was purified by silica gel column chromatography, using100:1→100:2/CH2Cl₂:95% EtOH as eluent, giving XLIV a white solid (619 mg79%).

[0241]¹H NMR (CDCl₃) δ0.87 (s, 9H) 0.97 (s, 9H) 2.13 (s, 3H) 4.87 (d,2H) 4.62 (s, 2H) 5.05 (br, 1H) 6.15 (br, 1H) 7.33 (m, 20H) 8.19 (s, 1H).¹⁹F NMR CDCl₃δ−120.40 (s).

[0242] To a solution of XLIV (391 mg. 0.5 mmol) in dry THF (10 ml) wasadded TBAF (0.32 ml, 2.2 mmol) and stirred at room temperature for 16hours. Removal of the solvent followed by purification by silica gelcolumn chromatography using 20:1→10:1/CN₂Cl₂: MeOH, as eluent gave XLVas a white solid. (135 mg, 89%).

[0243]¹H NMR (MeOD) δ2.40 (a, 3H) 4.39 (s, 2H) 4.94 (s, 2H) 6.98 (m, 2H)7.34 (m, 2M) 8.78 (s, 1H). ¹⁹F NMR (MeOD) δ−123.51 (m).

[0244] The syntheses of compounds XLVI aid XLVII followed the procedureoutlined for XLIV and XLV using 4-fluorophenylthioisocyanate in place of4-fluorophenylisocyanate. Yields obtained were 65% for XLVI and 60% forXLVII.

[0245]¹H NMR (XLVI) (CDCl₃) δ0.82 (s, 9H) 0.88 (s, 9H) 2.09 (s, 3H) 4.52(s, 2H) 5.00 (d, 2H 5.29 (s, 1H) 6.36 (t, 1H) 7.33 (m, 20H) 8.02 (s,1H). ¹⁹F NMR (CDCl₃) δ−113.42 (s).

[0246]¹H NMR (XLVII) (MeOD) δ2.41 (s, 3H) 3.07 (m. 1H) 3.54 (m, 1H) 4.91(s, 2H) 7.09 (t, 2H) 7.32 (q, 2H) 7.89 (s, 1H). ¹⁰F NMR (MeOD) δ −118.80(m).

Example 20

[0247] In Vivo Assay—Coronary Artery Ligation

[0248] Myocardial infarction was produced in male Sprague-Dawley rats(300-400 g) by occlusion of the left coronary artery. The rats werehoused in clear cages at room temperature and humidity on a 12 hourlight-dark cycle. Food and water wire supplied ad libitum. Rats wereanaesthized and The sit was incised along the left sternal border andthe fourth rib was cut proximal to the sternum and a retractor wasinserted. Me pericardial Sao was opened and the heart externalized. Theleft anterior descending coronary artery was ligated approximately 2 mmfrom its origin on the aorta using a 6-0 silk suture. The heart was thenrepositioned in the chest and the incision closed via purse-stringsutures. Sham-operated rats underwent substantially identical treatmentexcept that the artery was not ligated. All animals were allowed torecover, receiving food and water ad libitum for 21 days. Hemodynamicand histological assessments were made.

[0249] Occlusion of tie coronary artery rats has been shown to producemyocardial cell damage, which results in scar formation in the leftventricle and heart dysfunction. While the complete healing of the scartypically occurs within 3 weeks of the coronary occlusion, mild,moderate and severe stages of congestive heart failure have beenreported to occur at 4, 8 and 16 weeks after ligation. Accordingly, thecontractile dysfunction seen at 3 weeks after coronary occlusion in ratsis due to acute ischemic changes.

[0250] Rats were divided at random into five groups: sham operated,coronary artery ligated (untreated), coronary artery ligatedpyridoxal-5′-phosphate (PLP) treatment), and coronary artery ligated(compound XXIIb treated). Treatment wit PLP and compound XXIIb began 1hour after coronary occlusion (or sham operation), and continued for 21days. PLP or compound XXIIb (10 mg/kg) were administered daily (9 AM) bygastric tube. This dosage was chosen based on previous experience withPLP.

[0251] Mortality in all groups occurred only within the first 24 h aftercoronary ligation. While in the untreated group 50% of the rats died,the mortality rate was significantly reduced in the treated groups.

Example 21

[0252] In Vivo—Hemodynamic Changes

[0253] The animals prepared as described in Example 20 wereanaesthetized on the 21^(st) day with an injected cocktail of ketaminehydrochloride and xylazine. To maintain adequate ventilation, thetrachea was intubated. The right carotid artery was exposed and amicrochip pressure transducer was introduced (Model SPR-249, Millar,Houston, Tex.) into the left ventricle. The catheter was secured with asilk ligature around the artery, and various hemodynamic parametersincluding left ventricular systolic pressure (LVSP), left ventricularend diastolic pressure (LVEDP), rate of contraction (+dP/dt), and rateof relaxation (−dP/dt), were recorded and calculated with Acknowledge3.1 software (Biopac Systems Inc.). The animals were allowed 5-10minutes to stabilize, after which parameters were measured as averagesover three readings.

[0254] Average +dP/dt and −dP/dt values were reduced in the untreatedgroup compared to the sham control group. The experimental groupsreceiving P5P or compound XXIIb experienced recoveries in +dP/dt (rateof contraction) and −dP/dt (rate of relaxation) values.

[0255] LVSP (left ventricular systolic pressure) was decreased in theuntreated group compared to the sham control group, after 21 days ofcoronary ligation. Average LVEDP was increased in the untreated group,compared to the sham control group. Treatment with PLP, or compoundXXIIb yielded similar reduced rises in LVEDP in response to coronaryocclusion.

Example 22

[0256] In Vivo—Infarct Size and Scar Mass

[0257] After 21 days, once the hemodynamic data were obtained, theanimals were sacrificed and both average dry scar mass to left ventriclemass (n=5/group) and infarct size (n=5/group) were measured. For infarctsize measurement, hearts from the untreated and PLP/compoundXXIIb-treated groups were fixed in formalin and embedded in paraffin.Six evenly spaced slices were cut across the left ventricle. 5 μMsection were cut from each slice and mounted. Sections were stained withTrichrome to discriminate between fibrous scar and noninfarcted tissue.Using the free-drawing line tool in Scion Image v4.02b, infarct internalperimeter and left ventricle internal perimeter lengths were traced peroculum for each section. Noteworthy transmural scars were taken intoaccount. Infarct size was then expressed as the average scarperimeter/ventricle perimeter ratio. Scar mass measurements wereobtained by drying excised scar tissue and left ventricles at 50-60° C.for 72 hours.

[0258] The PLP-treated group had a reduced dry scar to left ventriclemass ratio. The compound XXIIb-treated group had scar/ventricle ratioreductions similar in magnitude to that seen in the PLP group. In theuntreated control group, average infarct size (as a percentage of leftventricle size) was about 45%; in the PLP and compound XIIb groups. theinfarct size was reduced.

Example 23

[0259] In Vivo—Hypertrophy

[0260] Hypertrophy is a physiological condition of enlargement(increased mass) due to increased stress. Cardiac hypertrophy isassessed by calculating the heart to body mass ratio. Treatment with PLPor compound XXIIb results in a decrease in cardiac hypertrophy, in therat model described in Example 20.

Example 24

[0261] Inhibition of Platelet Aggregation

[0262] Platelet rich plasma (PRP) was obtained by drawing whole bloodinto sodium citrate tubes (3.2%) and centrifuging at 700 rpm for 10minutes. Platelet poor plasma (PPP) was obtained by centrifuging theremainder of the sample util the platelets were removed (3200 rpm for 10minutes). The PRP was adjusted to a count of 280×10⁹/L using a mixtureof PRP and PPP. The incubation mixture consisted of 200 μL of plateletsand 25 μL of the appropriate compound (18 mM stock for a finalconcentration of 2 mM and 4.5 mM stock for a final concentration of 500μM), rendering an approximate final platelet count in the incubationmixture of 250×10⁹/L. After incubation (30 minutes at room temperature,the cuvettes were incubated for 3 minutes at 37° C., transferred to themixing wells and after the baselines transmittances were measured in anaggregometer (Chrono-Log 4), 25 μL of agonist was added to give a finalconcentration of 4 μM ADP, 1 μg/mL collagen, 5 μg/mL collagen or 12 μMthrombin receptor activating peptide (TRAP). Final transmittances werethen measured in the aggregometer (lower percent transmittancerepresents higher aggregation). Agonists concentrations were chosenbased on previous experience that indicated that these were the smallestconcentrations that would give the full of aggregation in tile normalpopulation Table 24-1 gives the results of the extent of aggregation forseveral compounds as percentage amplitude read directly from theaggregometer. TABLE 24-1 Extent of aggregation as a percentageamplitude, read directly from the aggregometer. Compound 5 μg/L 1 μg/mL12 μM Diluent of Tested collagen collagen 4 μM ADP TRAP Compound Saline82% 87% 83% 93% Control DMSO 85% 86% 55% 88% Control XL  5%  4%  4%  4%saline 2 Mm XL 52%  4%  9%  3% saline 500 μM XLVII 49% 30% 31% 42% 20%DMSO 2 mM in saline XXXV 67% 79% 15% 11% saline 2 mM XXXV 75% 23% 70%90% saline 500 μM XXIIb 77% 32% 46% 84% 1 part 2 mM DMSO 1 part salineXXIIb 82% 19% 59% 91% 1 part 500 μM DMSO 1 part saline PLP 89% 84% 16%91% saIine 2 mM PLP 87% 89% 69% 90% saline 500 μM

Example 25

[0263] In Vivo—Neuroprotection in Rat Embolic Stroke Model

[0264] Compound XL (see example 18, scheme 12) was assayed in a ratembolic stroke model.

[0265] Male Wistar rats, weighting 300-350 g, were purchased fromCharles River, St. Constant, Canada. Embolic focal cerebral ischemia wasinduced by embolizing a pre-formed clot into the middle cerebral artery(MCA). (For details of the procedure see Wang CX, Yang Y, Shuib A. Afocal embolic model of cerebral ischemia in rats. Brain Res. Protoc.7:115-120, 2001. Wang C, Yang T and Shuaib A: An improved version ofembolic model of brain ischemic injury in the rat. J. Neuzosci. Meth.109:147-151, 2001).

[0266] The Study consisted of four groups (n=10). Animals were randomlyassigned to one of the four groups. Treatments were started at 1 h afterMCA occlusion. XL was dissolved in 0.5% DMSO (1.5 ml) and infused over aperiod of 60 min i.v. in the experimental groups, XL, 20, 40 or 80 mg/kgwas infused. In the control group, 0.5% DMSO (1.5 ml) was infused.

[0267] The procedures for assessment of infarct volume were detailedpreviously (Bederson J B, Pitts L H, Tsuji M, Nishimura M C, Davis R L,Bartcowski H. Rat middle cerebral artery occlusion: evaluation of themodel and development of a neurologic examination. Stroke 17:472-476.1986. Yang, Y., Shuaib, A., and Li, Q. Quantification of infarct size onfocal cerebral ischemia model of rats using a simple and economicalmethod. J. Neurosci. Meth., 84:9-16, 1998). Briefly, at 48 h afterischemia the anesthetized animal was sacrificed by decapitation and thebrain, removed. The volume of infarction was measured in the brainsections stained with 2,3,5-triphenyltetrazolium chloride (TTC) solutionwith following formula, corrected infarct size=the volume of the lefthemisphere —(the volume of the right hemisphere-measured infarctvolume). The percentage of infarct volume was obtained by calculatingthe portion of the corrected infarct volume and dividing it by the totalvolume of the left hemisphere. Brain swelling was determined using aformula: swelling (edema)=(volume of the right hemisphere=volume of theleft hemisphere)/volume of the left hemisphere. Hemorrhagetransformation was identified from the TTC stain brain Sections.

[0268] Neurological deficits and seizure activities were recorded at 2,24 and 48 hours after embolization. Neurological deficits weredetermined using a modified Bederson's scoring system (Bederson J R,Pitts L K Tsuji M, Nishimura M C, Davis R L, Bartkowski H. Rat middlecerebral artery occlusion: evaluation of the model and development of aneurologic examination. Stroke 17:472476, 1986). 0: no observabledeficit. 1: forelimb flexion; 2: forelimb flexion plus decreasedresistance to lateral push; 3: unidirectional circling; 4:unidirectional circling plus decreased level of consciousness. Seizureactivity, hemorrhage and mortality were also recorded. Seizure was alsorecorded. Results are shown in Table 25-1

[0269] The differences of infarction volume were analyzed with one wayANOVA followed by Dunnett's test. Neurological deficit scores werereported as medians and interquartile ranges, considering 25-75^(th)percentile. The neurological scores were analyzed with Kruskal-Wallistest when more than two groups, and analyzed with Mann-Whitney test whencomparison was done in two groups. The rates of mortality and seizureoccurrence following different treatments were compared with Chi-squaretest. Differences were considered significant when p<0.05.

[0270] Infarction volume and brain edema. Embolizing a pre-formedthrombus resulted in an infarction in the territory irrigated by MCA,mainly located in the cerebral cortex and striatum. In the controlgroup, the infarction volume was 40±6% (mean±SEM) at 48 h after theembolization. In the groups administrated XL 10, 20 or 40 mg/kg at 1 hafter embolization, the infarction volumes were 30.5±4.8, 26.5±4.4 and25.94±3.4% respectively, which was significantly different from thecontrol group. Ischemic brain edema was 5.7±1.3, 6.8±1.9, 5.9±1.3 and7.9±2.1 respectively. There was no significant difference among thegroups.

[0271] Neurological deficits. Neurological deficits are shown in Table25-1. In all four groups, the Bederson score was 3 at 2 after theischemic injury. Kruskal-Wallis test analysis showed that there weresignificant differences among the groups at 48 h but not at 24 h.Compared to the score recorded at 2 h after the injury, it did notsignificantly improve at either 24 h or 48 h after the injury in thecontrol group. In the rats receiving the drug injection of 10, 20 or 40mg/kg at 2 h after the injury, neurological scores improvedsignificantly at 24 h after the injury when compared with that at 2 hafter the injury (P<005). Neurological scores also improvedsignificantly at 48 h after the injury when compared with that at 2 hafter the injury (P<0.05).

[0272] Seizure occurrence. Seizure occurred in 2 rats in the controlgroup, 3 rats in the 10 mg/kg group. 2rats in the 20 mg/kg group and 2rats in the 40 mg/kg group.

[0273] Hemorrhage rate. Hemorrhage was observed in 1 rats in the controlgroup, 2 rats in the 10 mg/kg group, 1 rat in the 20 mg/kg group and 2rats in the 40 mg/kg group.

[0274] Mortality rate. Pro-maturely death occurred 1 rat in controlgroup, 1 in the 10 mg/kg group and 1 in the 20 mg/kg group. TABLE 25-1Neurological deficits in rats received different doses of XL afterinjury^(a) Saline 10 mg^(b) 20 mg 40 mg Group n = 10 n = 10 n = 10 n =10  2 h 3 (3-3) 3 (3-3) 3 (3-3) 3 (3-3) 24 h 3 (3-3) 2 (2-3)* 2 (2-3)* 2(2-3)* 48 h 3 (3-3) 2 (2-3)* 2 (2-3)* 2.5 (2-2)*

Example 26

[0275] In Vivo—Assay in Rat Cardiac Ischemia Reperfusion Model

[0276] Compound XL (see example 18, scheme 12) was assayed in a ratcardiac ischemia reperfusion model

[0277] Seventy-nine male Wistar rats (200-300 g, Tuck, Rayleigh, Essex,U.K.) were anesthetized with thiopentone sodium (Intraval®, 120 mg/kgi.p.; Rhone-Memrieux, Essex, U.K.). The rats were tracheotomized,intubated and ventilated with a Harvard ventilator (70 strokes/min,tidal volume: 8-10 ml/kg, inspiratory oxygen concentration: 30%). Bodytemperature was maintained at 38±1° C. The right carotid artery wascannulated and connected to a pressure transducer (Spectramed, P23XL) tomonitor mean aterial blood pressure (MAP) and head rate (HR), which werecontinuously recorded on a 4-channel Grass 7D polygraph recorder (Grass,Mass., U.S.A.). The right jugular vein was cannulated for theadministration of drugs. Subsequently, a lateral thoracotomy wasperformed and the heart was suspended in a temporary pericardial cradle.A snare occluder was placed around the left anterior descending coronaryartery (LAD). After completion of the surgical procedure the animalswere allowed to stabilize for 30 min before LAD ligation. The coronaryartery was occluded at time 0 by tightening of the occluder. After 25min of acute myocardial ischemia, the occluder was re-opened to allowthe reperfusion for 2 h. Following the 2 h reperfusion period, thecoronary artery was re-occluded and Evans Blue dye (1 ml of 2% w/v) wasinjected into the left ventricle, via the right carotid artery cannula,to distinguish between perfused and non-perfused area at risk (AR)sections of the heart. The Evans Blue solution stains the perfusedmyocardium, while the occluded vascular bed remains uncolored. Theanimals were killed with an overdose of anesthetic and the heartexcised. It was sectioned into slices of 3-4 mm, the right ventricularwall was removed, and the AR (pink) was separated from the non-ischemic(blue) area. The AR was cut into small pieces and incubated withp-nitroblue tetrazolium (NBT, 0.5 mg/ml) for 40 min at 37° C. In thepresence of intact dehydrogenase enzyme systems (viable myocardium),nitro-blue tetrazolium (NBT) forms a dark blue formazan, whilst areas ofnecrosis lack deydogmenase activity and therefore fall to stain. Pieceswere separated according to staining and weighed to determine theinfarct size as a percentage of the weight of the AR.

[0278] The following experimental groups were studied:

[0279] (1) Sham-operation (no occlusion of the LAD) plus administrationof saline (Sham-Saline, 1 ml/kg i.v. n=9).

[0280] (2) LAD-occlusion (25 min) and reperfusion (2 h) plusadministration of saline (myocardial infarction (“MI”)-Saline, 1 ml/kgiv. bolus, 5 min prior to LAD occlusion) (n=10).

[0281] (3) LAD-occlusion and reperfusion plus administration of XL (25mg/kg i.v. bolus at 5 min prior to LAD occlusion) (n=10).

[0282] All data were expressed as mean±i s.e.mean of n observations,where n represented the number of animals studied. Infarct size wasanalyzed by 1-factorial ANOVA, followed by a Bonferroni's post hoc testfor multiple comparisons. Hemodynamic data were analyzed by 2-way ANOVAfollowed by a Bonferroni's test. A P-value of less than 0.05 wasconsidered to be statistically significant (when compared to MI-Saline).

[0283] The baseline values of mean arterial pressure (MAP) in all groupsof animals were not significantly different between groups (P>0.05,Table 26-1). The MAP of rats subjected to LAD occlusion wassignificantly lower at 15 min of regional myocardial ischemia (P<0.05.when compared to sham operated animals, Table 26-1). In addition therewas a progressive decline in MAP during the reperfusion period (P<0.05,Table 26-1). The fall in MAP observed in MI-rats treated with the testcompound XL (25 mg/kg i.v.)] was similar to the fall in MAP observed inthe rats subjected to myocardial ischemia and treated with Saline.(P>0.05, Table 26-1).

[0284] Baseline values of heart rate (ER) in all groups of animals werenot significantly different between groups (P>0.05, Table 26-1). In ratssubjected to myocardial ischemia and reperfusion, there was no change inHR. The test compound had no significant effect on HR in rats subjectedto regional myocardial ischemia and reperfusion (P>0.05, Table 26-1).

[0285] Baseline values of the power ratio index (PRI) in all groups ofanimals were not significantly different between groups (P>0.05, Table26-1). In rats subjected to myocardial ischemia and reperfusion, therewas a progressive fall in PRI (P>0.05, Table 26-1). The fall in PRIobserved in MI-rats treated with the test compound XL (25 mg/kg i.v.)]was similar to the fall in PRI observed in the MI-rat treated withSaline. (P>0.05. Table 26-1).

[0286] The mean values for the area at risk of infarction (AR) rangedfrom 46±3% to 58±3% and were similar in all animal groups studied(P>0.05, FIG. 1a). In rats, which were treated with saline (vehicle),occlusion of the LAD (for 25 min) followed by reperfusion (for 2 h)resulted in an infarct size of 48±3% of the AR (MI-Saline, n=10).

[0287] Intravenous administration of XL (25 mg/kg i.v. bolus, 5 minutesbefore the onset of regional ischemia) resulted in a significantreduction in infarct size (P<0.05, FIG. 1b).

[0288] Thus intravenous administration of the test compound XL resultsin a significant reduction in the infarct size caused by regionalmyocardial ischemia and reperfusion in an anesthetized mammalianpatient. TABLE 26-1 Table illustrating the alterations in hemodynamicparameters throughout the experiment; 25 mins LAD occlusion, 2 hreperfusion Occlusion (mins) Reperfusion (mins) n Baseline 15 25 60 120Sham Saline 9 MAP 147 ± 5 149 ± 9* 146 ± 9 133 ± 9* 122 ± 5* HR 421 ± 10424 ± 13 423 ± 14 410 ± 11 403 ± 11 PRI  62 ± 2  36 ± 14  61 ± 4  55 ± 4 48 ± 2 MI Saline 10 MAP 134 ± 6 119 ± 9 117 ± 9  87 ± 9  91 ± 6 1 ml/kgsaline HR 427 ± 11 427 ± 13 423 ± 10 401 ± 12 397 ± 9 t = −5 mins PRI 57 ± 3  52 ± 5  51 ± 4  35 ± 4  36 ± 2 XL 8 MAP 121 ± 67 119 ± 6 118 ±6  89 ± 8  70 ± 7 25 mg/kg i.v.; HR 443 ± 9 435 ± 11 460 ± 10 412 ± 10414 ± 13 B PRI  54 ± 3  52 ± 3  55 ± 3  36 ± 3  29 ± 3 t = −5 mins

Example 27

[0289] Inhibition of Platelet Aggregation

[0290] XL was capable of inhibiting the aggregation of platelets drawnfrom patents known to have heparin induced thrombocytopenia (HIT).Platelets from HIT patients were challenged with 0.5 U/ml concentrationof heparin in the presence and absence of drug. At 500 micromolar XL wasable to completely suppress the aggregation. XL is useful as anantiplatelet treatment for HIT patients who cannot be administeredheparin due to the immune reaction with heparin that leads toaggregation rather than thrombolysis.

Example 28

[0291] Treatment of Heparin Induced Thrombocytopenia (HIT)

[0292] XL inhibited the effects of heparin induced thrombocytopenia(HIT). As shown in Table 28-1,500 micromolar XL was able to completelysuppress the affects of HIT. TABLE 28-1 Amplitude of ATP release Ratioof spike (low Low/High Aggregation Concentration Heparin) Heparin (LowHeparin) Patient 1 0 363 42.4 Present 500 μM 14 1.9 Absent  2 mM 28 2.7Absent Patient 2 0 286 37.6 Present 500 μM 11 1.6 Absent  2 mM 12 1.6Absent

[0293] Platelets were drawn from HIT patients and challenged withheparin to a final concentration of 0.5 U/ml of low molecular weightheparin, in the presence and absence of the drug. ATP is released andaggregation occurs when patients exhibit HIT. ATP was detected in a lumiaggregometer (Chronolog Corp). A luciferin-luciferase mixture was usedto detect ATP release. The luciferin-luciferase mixture was previouslycalibrated to a known amount of ATP. Thus a spike in ATP releaseindicated platelet aggregation. As shown by the data, HIT patents have alarge amplitude release when there is no XL compound present. When XLcompound is present it inhibits this characteristic release of ATP.

[0294] Low heparin is low molecular weight heparin and high heparin isunfractionated heparin. The presence of low molecular weight heparinwill cause platelet aggregation. All samples were treated with aconstant final concentration of heparin; 0.5U/ml-low and 100U/ml high.HIT causes platelet aggregation in the Low Concentration of Heparinonly, as well as a ration of >5.0 when comparing the Low/Highconcentration of Heparin. Thus, the data presented in Table 28-1 showsthat XL is effective as an treatment for HIT patients whom otherwisecannot be administered heparin due to the immune reaction with heparinthat leads to aggregation rather than thrombolysis.

[0295] It should be noted that, as used in this specification and theappended claims, the singular forms “a,” “an,” and “the” include pluralreferents unless the content clearly dictates otherwise. Thus forexample, reference to a composition containing “a compound” includes amixture of two or more compounds.

[0296] Although embodiments of the invention have been described above,it is not limited thereto, and it will be apparent to persons skilled inthe art that numerous modifications and variations form part of thepresent invention insofar as they do not depart from the spirit, nature,and scope of the claimed and described invention.

We claim:
 1. A method of treating or preventing heparin induced thrombocytopenia (HIT), in a mammalian patient comprising administering a therapeutically effective amount of a compound of Formula V:

or a pharmaceutically acceptable acid addition salt thereof, wherein: R₅ is CH₂OH or CHO; R₁ is

n is an integer of 1 to 5; R₂, R₃, and R₄ are each independently hydrogen; alkyl; aryl or biaryl, wherein the aryl or biaryl can be substituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy, amino; acylamino; anilino, wherein the aniline ring can be substituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy; nitro; or guanidino.
 2. The method of claim 1 wherein said compound is administered to induce a plasma level of said compound of between about 100 nM and 10,000 μM
 3. The method of claim 2 wherein said plasma level is between 200 μM and 4000 μM.
 4. The method of claim 3 wherein said plasma level is between 500 μM and 2000 μM
 5. The method of claim 1 wherein said therapeutically effective dose is between about 0.1 to 100 mg/kg body weight per day.
 6. The method of claim 5 wherein said compound is administered intravenously, orally, sublingually, intraperitoneally, transdermally, intramuscularly or subcutaneously.
 7. The method of claim 1 wherein said compound is administered intravenously.
 8. The method of claim 1 wherein said compound is administered orally.
 9. The method of claim 1 wherein the compound is


10. The method of claim 3 wherein the compound is


11. A method of treating or preventing stroke in a mammalian patient comprising administering a therapeutically effective amount of a compound of Formula V:

or a pharmaceutically acceptable acid addition salt thereof wherein: R₅ is CH₂OH or CHO; R₁ is

n is an integer of 1 to 5; R₂, R₃, and R₄ are each independently hydrogen; alkyl; aryl or biaryl, wherein the aryl or biaryl can be substituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy; amino; acylamino; anilino, wherein the aniline ring can be substituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy; nitro; or guanidino.
 12. The method of claim 11 wherein said compound is administered to induce a plasma level of said compound of between about 100 nM and 10,000 μM.
 13. The method of claim 11 wherein said therapeutically effective dose is between about 0.1 to 100 mg/kg body weight per day.
 14. The method of claim 13 wherein said dosage is between 1 mg/kg and 100 mg body weight.
 15. The method of claim 14 wherein said dosage is between 5 mg/kg sand 80 mg/kg body weight.
 16. The method of claim 13 wherein said compound is administered intravenously, orally, sublingually, intraperitoneally, transdermally, intramuscularly or subcutaneously.
 17. The method of claim 11 wherein said compound is administered intravenously.
 18. The method of claim 11 wherein said compound is administered orally.
 19. The method of claim 11 wherein the compound is


20. The method of claim 13 wherein the compound is


21. A method of treating or preventing ischemia in a mammalian patient comprising administering a therapeutically effective amount of a compound of Formula V:

or a pharmaceutically acceptable acid addition salt thereof, wherein: R₅ is CH₂OH or CHO; R₁ is

n is an integer of 1 to 5; R₂, R₃, and R₄ are each independently hydrogen; alkyl; aryl or biaryl, wherein the aryl or biaryl can be substituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy; amino; acylamino; anilino, wherein the aniline ring call be substituted with a cyano, alky alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy; nitro; or guanidino.
 22. The method of claim 21 wherein said compound is administered to induce a plasma level of said compound of between about 100 nM and 10,000 μM.
 23. The method of claim 21 wherein said therapeutically effective dose is between about 0.1 to 100 mg/kg body weight per day.
 24. The method of claim 23 wherein said dosage is between 1 mg/kg and 100 mg/kg body weight.
 25. The method of claim 24 wherein said dosage is between 5 mg/kg and 80 mg/kg body weight.
 26. The method of claim 23 wherein said compound is administered intravenously, orally, sublingually, intraperitoneally, transdermally, intramuscularly or subcutaneously.
 27. The method of claim 21 wherein said compound is administered intravenously.
 28. The method of claim 21 wherein said compound is administered orally.
 29. The method of claim 21 wherein the compound is


30. The method of claim 23 wherein the compound is


31. A method of providing a neuroprotective effect in a mammalian patient comprising administering a therapeutically effective amount of a compound of Formula V:

or a pharmaceutically acceptable acid addition salt thereof, wherein: R₅ CH₂OH or CHO; R₁ is

n is an integer of 1 to 5; R₂, R₃, and R₄ are each independently hydrogen; alkyl; aryl or biaryl. wherein the aryl or biaryl can be substituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy, amino; acylamino; anilino, wherein the aniline ring can be substituted with a cyano, alkyl, alkoxy, amino, hydroxy, halo, nitro, or alkanoyloxy; nitro; or guanidino.
 32. The method of claim 31 wherein said compound is administered to in a dosage of between 0.1 mg/kg and 100 mg/kg.
 33. The method of claim 32 wherein said dosage is between 1 mg/kg and 100 mg/kg.
 34. The method of claim 33 wherein said plasma level is between 5 mg/kg and 80 mg/kg
 35. The method of claim 31 wherein said compound is administered to induce a plasma level of said compound of between about 100 nM and 10,000 μM.
 36. The method of claim 33 wherein said compound is administered intravenously, orally, sublingually, intraperitoneally, transdermally, intramuscularly or subcutaneously.
 37. The method of claim 31 wherein said compound is administered intravenously.
 38. The method of claim 31 wherein said compound is administered orally.
 39. The method of claim 31 wherein the compound is


40. The method of claim 33 wherein the compound is 